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[KD Validated] eIF4E Rabbit mAb (A25608)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat, Monkey

Western blot analysis of lysates from wild type (WT) and eIF4E knockdown (KD) 293T cells using [KD Validated] eIF4E Rabbit mAb (A25608) at 1:3000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 1s.

Western blot analysis of various lysates using [KD Validated] eIF4E Rabbit mAb (A25608) at 1:3000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

Western blot analysis of various lysates using [KD Validated] eIF4E Rabbit mAb (A25608) at 1:3000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

ABclonal:Immunofluorescence - [KD Validated] eIF4E Rabbit mAb (A25608)

Confocal imaging of 293T cells using [KD Validated] eIF4E Rabbit mAb (A25608, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of Hep G2 cells using [KD Validated] eIF4E Rabbit mAb (A25608, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of NIH/3T3 cells using [KD Validated] eIF4E Rabbit mAb (A25608, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of PC-12 cells using [KD Validated] eIF4E Rabbit mAb (A25608, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

ABclonal:Immunoprecipitation - [KD Validated] eIF4E Rabbit mAb (A25608)

Immunoprecipitation of [KD Validated] eIF4E in 200 µg extracts from 293T cells using 0.5 µg [KD Validated] eIF4E Rabbit mAb (A25608). Western blot analysis was performed using [KD Validated] eIF4E Rabbit mAb (A25608) at 1:3000 dilution.

Overview

Product name	[KD Validated] eIF4E Rabbit mAb
Catalog No.	A25608
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC66295

Background

The protein encoded by this gene is a component of the eukaryotic translation initiation factor 4F complex, which recognizes the 7-methylguanosine cap structure at the 5' end of messenger RNAs. The encoded protein aids in translation initiation by recruiting ribosomes to the 5'-cap structure. Association of this protein with the 4F complex is the rate-limiting step in translation initiation. This gene acts as a proto-oncogene, and its expression and activation is associated with transformation and tumorigenesis. Several pseudogenes of this gene are found on other chromosomes. Alternative splicing results in multiple transcript variants.

Immunogen information

Immunogen	A synthetic peptide corresponding to a sequence within amino acids 1-100 of human eIF4E (NP_001959.1).
Sequence	MATVEPETTPTPNPPTTEEEKTESNQEVANPEHYIKHPLQNRWALWFFKNDKSKTWQANLRLISKFDTVEDFWALYNHIQLSSNLMPGCDYSLFKDGIEP
Gene ID
Swiss Prot
Synonyms	CBP; EIF4F; AUTS19; EIF4E1; eIF-4E; EIF4EL1
Calculated MW	25kDa/27kDa/29kDa
Observed MW	28kDa/

Applications

Reactivity	Human, Mouse, Rat, Monkey
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:3000 - 1:18000 IF/ICC 1:200 - 1:2000 IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunofluorescence Immunoprecipitation
Positive samples	293T, HeLa, COS-7, NIH/3T3, RAW 264.7
Cellular location	Cytoplasm, P-body, Stress granule, Cytosol, Nucleus, P-body, chromatoid body, cytoplasm, cytoplasmic ribonucleoprotein granule, cytoplasmic stress granule, cytosol, eukaryotic translation initiation factor 4F complex, extracellular exosome, glutamatergic synapse, nucleus, perinuclear region of cytoplasm, postsynaptic cytosol.